A method for increasing the yield of riboflavin by using an oxygen carrier and its fermentation device

A technology of riboflavin and oxygen carrier, applied in specific-purpose bioreactors/fermenters, methods based on microorganisms, fermentation, etc., can solve the complex reaction of pure chemical synthesis, affect the prospects of industrial production, and riboflavin production Reduce and other problems, to achieve the effect of improving the substrate conversion rate, shortening the fermentation cycle, and increasing the dissolved oxygen capacity

Active Publication Date: 2021-07-27
HEBEI SHENGXUE DACHENG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the large-scale synthesis of riboflavin is mainly divided into chemical synthesis and microbial fermentation. However, the reaction of pure chemical synthesis is too complicated, the cost is high and the yield is low, which greatly affects the prospect of industrial production.
[0004] The technology of riboflavin fermented by Bacillus subtilis has become relatively mature, replacing the previous complicated and inefficient chemical synthesis methods, but there are still some problems in the fermentation process. High, especially in the later stage of fermentation, the oxygen consumption of fermentation is high, once the dissolved oxygen is lower than a certain level, the riboflavin production will be greatly reduced
[0005] For a long time, most of the research on riboflavin production has been the transformation of genetically engineered bacteria, which is costly, time-consuming, and has a low success rate. The impact on production and cost caused by the oxygen consumption of the bacteria itself has been ignored. Therefore, how to meet the requirements during the fermentation process? The oxygen demand of Bacillus subtilis, increasing the production of riboflavin and reducing the cost are difficult problems that need to be solved at present. Therefore, we propose a method of using oxygen carrier to increase the production of riboflavin and its fermentation device to solve the above problems

Method used

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  • A method for increasing the yield of riboflavin by using an oxygen carrier and its fermentation device
  • A method for increasing the yield of riboflavin by using an oxygen carrier and its fermentation device
  • A method for increasing the yield of riboflavin by using an oxygen carrier and its fermentation device

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Experimental program
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Embodiment 1

[0038] This embodiment includes the following steps:

[0039] Seed culture: Inoculate the riboflavin-producing bacteria Bacillus subtilis into the seed medium, and culture it with shaking at 35°C and 200-240rpm on a fermentation device for 24 hours to obtain the seed medium;

[0040] Fermentation culture: Inoculate the obtained seed culture liquid into the fermentation medium, add sterile oxygen carrier n-dodecane at 0 hours after the fermentation starts, and the amount of n-dodecane added is 1%, and finally obtain the fermentation liquid containing riboflavin , the fermentation condition is: 500mL baffle Erlenmeyer flask, liquid volume is 30 milliliters, at 30-38 ℃, 200-240rpm fermenting equipment shaking culture for 120 hours.

[0041] Yield detection: the method is the same as that of Comparative Example, and the content of riboflavin in the fermented liquid of Example 1 is 12632 μg / mL, which is 10.6% higher than that of Comparative Example.

Embodiment 2

[0043] This embodiment includes the following steps:

[0044] Seed culture: Inoculate the riboflavin-producing bacteria Bacillus subtilis into the seed medium, and culture it with shaking at 35°C and 200-240rpm on a fermentation device for 24 hours to obtain the seed medium;

[0045] Fermentation culture: Inoculate the obtained seed culture liquid into the fermentation medium, add sterile oxygen carrier n-dodecane to the fermentation medium for 60 hours, and the addition amount of n-dodecane is 4%, finally obtain the fermentation liquid containing riboflavin, The fermentation conditions are: 500mL baffled Erlenmeyer flask with a liquid volume of 30ml, shake culture at 30-38°C, 200-240rpm fermentation equipment for 90 hours.

[0046] Yield detection: the method is the same as that of the comparative example. The riboflavin content in the fermented liquid of Example 2 is 13034 μg / mL, which is 12.4% higher than that of the comparative example, and the fermentation period is short...

Embodiment 3

[0048] This embodiment includes the following steps:

[0049] Seed culture: Inoculate the riboflavin-producing bacteria Bacillus subtilis into the seed medium, and culture it with shaking at 35°C and 200-240rpm on a fermentation device for 24 hours to obtain the seed medium;

[0050] Fermentation culture: inoculate the obtained seed culture solution into the fermentation medium, add sterile oxygen carrier n-dodecane to the fermentation medium for 50 hours, the addition of n-dodecane is 3%, and finally obtain the fermentation solution containing riboflavin, The fermentation conditions are: 500mL baffled Erlenmeyer flask with a liquid volume of 30ml, shake culture at 30-38°C, 200-240rpm on a fermentation device for 105 hours.

[0051] Yield detection: the method is the same as that of the comparative example, and the riboflavin content in the fermented liquid of Example 3 is 15820 μg / mL, which is 38.5% higher than that of the comparative example, and the fermentation period is s...

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Abstract

The invention discloses a method for increasing the yield of riboflavin by using an oxygen carrier and a fermentation device thereof. S1, seed cultivation: inoculate the riboflavin-producing bacteria Bacillus subtilis into the seed culture medium, and cultivate to obtain the seed culture liquid; S2, fermentation culture: the obtained seed culture solution is inoculated into the fermentation medium, and then a sterile oxygen carrier is added to ferment to obtain riboflavin. In the present invention, by adding a suitable oxygen carrier in the culture medium and cultivating it in the fermentation device of the present invention, the dissolved oxygen concentration in the culture medium is improved simultaneously by chemical and physical methods, thereby promoting the riboflavin production of Bacillus subtilis The bacterial metabolism of Bacillus improves the substrate conversion rate, increases the synthesis amount of riboflavin from 11424 μg / mL to 15820 μg / mL, shortens the fermentation period and reduces the production cost.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation production, in particular to a method for increasing riboflavin production by using an oxygen carrier and a fermentation device thereof. Background technique [0002] Riboflavin is a kind of natural pigment soluble in water. It acts as a coenzyme to promote metabolism and exists in the form of flavin mononucleotide and flavin adenine dinucleotide in organisms. Riboflavin can be used to treat riboflavin deficiency, conjunctivitis, nutritional ulcers, and systemic nutritional disorders. Riboflavin lesions are mostly manifested as inflammation of the mouth, eyes and external genitalia, such as angular cheilitis, cheilitis, glossitis, conjunctivitis and scrotitis, etc. Therefore, riboflavin has a wide range of applications in food, feed, and pharmaceutical industries. application prospects. [0003] At present, the large-scale synthesis of riboflavin is mainly divided into chemical sy...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P25/00C12M1/38C12M1/36C12M1/34C12R1/125
CPCC12P25/00C12M23/48C12M41/12C12M41/34C12M23/46C12M27/16C12M29/00
Inventor 周景王绘砖魏伟林青曹建全
Owner HEBEI SHENGXUE DACHENG PHARMA
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